Packer Adam M, Russell Lloyd E, Dalgleish Henry W P, Häusser Michael
1] Wolfson Institute for Biomedical Research, University College London, London, UK. [2] Department of Neuroscience, Physiology and Pharmacology, University College London, London, UK.
Nat Methods. 2015 Feb;12(2):140-6. doi: 10.1038/nmeth.3217. Epub 2014 Dec 22.
We describe an all-optical strategy for simultaneously manipulating and recording the activity of multiple neurons with cellular resolution in vivo. We performed simultaneous two-photon optogenetic activation and calcium imaging by coexpression of a red-shifted opsin and a genetically encoded calcium indicator. A spatial light modulator allows tens of user-selected neurons to be targeted for spatiotemporally precise concurrent optogenetic activation, while simultaneous fast calcium imaging provides high-resolution network-wide readout of the manipulation with negligible optical cross-talk. Proof-of-principle experiments in mouse barrel cortex demonstrate interrogation of the same neuronal population during different behavioral states and targeting of neuronal ensembles based on their functional signature. This approach extends the optogenetic toolkit beyond the specificity obtained with genetic or viral approaches, enabling high-throughput, flexible and long-term optical interrogation of functionally defined neural circuits with single-cell and single-spike resolution in the mouse brain in vivo.
我们描述了一种全光学策略，用于在体内以细胞分辨率同时操纵和记录多个神经元的活动。我们通过共表达红移视蛋白和基因编码钙指示剂，进行了双光子光遗传学激活和钙成像同步操作。空间光调制器允许对数十个用户选择的神经元进行时空精确的并发光遗传学激活，同时快速钙成像同步提供了具有可忽略光学串扰的高分辨率全网络操纵读数。在小鼠桶状皮层进行的原理验证实验表明，可在不同行为状态下对同一神经元群体进行询问，并根据其功能特征对神经元集群进行靶向。这种方法将光遗传学工具包扩展到了超越遗传或病毒方法所能获得的特异性，能够在小鼠大脑体内以单细胞和单峰分辨率对功能定义的神经回路进行高通量、灵活且长期的光学询问。
